Cleavage of nucleic acids in a specific location is frequently used in many genetic engineering techniques. There are many methods to fragment DNA molecules, including the widely used commercially available restriction enzymes. Not many RNA processing enzymes are known and most of them are characterized by a low sequential specificity or its total absence.
WO 2010076939 A1 relates to compositions and methods for carrying out targeted genetic recombination or mutation using the chimeric zinc finger nuclease. WO 03087341 A2 relates to the use of a zinc finger nuclease for the targeted editing of the human cystic fibrosis transmembrane conductance regulator gene, thereby providing a potential therapy for cystic fibrosis. WO 2009146179 A1 relates to the development of a highly efficient and easy-to-practice modular-assembly method using publicly available zinc fingers to make zinc finger nucleases that are able to modify the DNA sequences of several genomic sites in human cells. From descriptions WO2007014181A2 and WO2007014182A2 there are known fusions of many zinc finger domains and Fold nucleases that facilitate targeted genome editing. However, unlike in the former case, the fusion of native proteins alone, RNase HI and the zinc finger, does not lead to a sequence-specific enzyme.
The method for determination of the sequence preference of DNA-RNA hybrid binding protein(s) or its domain(s) and by this determining the sequence recognized by a DNA-RNA hybrid binding protein is a modification of the SELEX procedure. SELEX stands for systematic evolution of ligands by exponential enrichment. The principle of this method is based on iterative selection and enrichment of molecules from a large diverged library of nucleic acids sequences that exhibit a high affinity towards a ligand. The enrichment step is accomplished by binding of the nucleic acids to a ligand and removal of the unbound sequences.
WO 2010076939 A1 relates to the compositions and methods for carrying out the targeted genetic recombination or mutation using the chimeric zinc finger nuclease. WO 03087341 A2 relates to the use of a zinc finger nuclease for the targeted editing of the human cystic fibrosis transmembrane conductance regulator gene, thereby providing a potential therapy for cystic fibrosis. WO2009146179 A1 relates to the development of a highly efficient and easy-to-practice modular-assembly method using publicly available zinc fingers to make zinc finger nucleases that are able to modify the DNA sequences of several genomic sites in human cells.
Herskovitz M. A. et al., Mol Microbiol. 2000 December: 38(5):1027-33, “Endoribonuclease RNase III is essential in Bacillus subtilis” relates to growth of a strain in which Bs-RNase III (rncS) expression was dependent upon transcription of rncS from a temperature-sensitive plasmid and at the non-permissive temperature resulted in 90-95% cell death, and virtually all the cells that survived retained the rncS-expressing plasmid. Thus, authors concluded that rncS is essential in B. subtilis. Dasgupta S. et al., Mol Microbiol. 1998; 28 (3): 629-40, “Genetic uncoupling of the dsRNA-binding and RNA cleavage activities of the Escherichia coli endoribonuclease RNase III—the effect of dsRNA binding on gene expression.” describes the phenotypes of bacteria carrying point mutations in mc, the gene encoding RNase III. Karen Shahbabian et al., The EMBO Journal (2009) 28, 3523-3533, “RNase Y, a novel endoribonuclease, initiates riboswitch turnover in Bacillus subtilis” describes an essential protein of earlier unknown function, YmdA, identified as a novel endoribonuclease (now called RNase Y) that was capable of preferential cleaving in vitro of the 5′-monophosphorylated yitJ riboswitch upstream of the SAM-binding aptamer domain.
US 2006057590 relates to the generation of a double stranded RNA molecule that substantially covers the whole transcribed region of a gene and cleaving this molecule using an RNA endonuclease to generate small RNA molecules, which are already or may be subsequently labeled. JP54059392 patent relates to a novel nuclease B-1 which attacks the single chain scission area of one of the DNA chains of a double-stranded deoxyribonucleic acid, and specifically splits the other DNA chain at its complementary area.
Citation or identification of any document in this application is not an admission that such document is available as prior art to the present invention.